Arylethanolamine derivatives and their use as agonists of atypical beta-adrenoceptors

ABSTRACT

The present invention relates to phenethanolamine derivatives of formula (I) wherein R 1  represents an aryl group optionally substituted by one or more substituents selected from halogen, hydroxy, C 1-6  -alkoxy, C 1-6  -alkyl, nitro, cyano, hydroxymethyl and trifluoromethyl; R 2  represents hydrogen or C 1-6  -alkyl; R 3  represents a group (A) where the ring is substituted by one to four further substituents selected from C 1-6  -alkyl, halogen, trifluoromethyl, and C 1-6  -alkoxy; or R 3  represents a group (B) where the aromatic ring is optionally substituted by up to three further substituents selected from C 1-6  -alkyl, halogen, trifluoromethyl, and C 1-6  -alkoxy; R 4  represents hydrogen, or C 1-6  -alkyl; R 5  represents ZCH 2  CO 2  H wherein Z represents a bond, or O; Y represents (CH 2 ) n  where n is 1-3; and physiologically acceptable derivatives thereof; to process for their preparation; and their use in the treatment of conditions susceptible of amelioration by an atypical beta-adrenoceptor agonist.

This application is filed pursuant to 35 U.S.C. §371 as a United StatesNational Phase Application of International Application No.PCT/EP96/05469 filed Dec. 6, 1996 which claims priority from GB9525177.3 filed Dec. 8, 1995.

This invention relates to a new class of chemical compounds and to theiruse in medicine. In particular, the invention concerns novelphenethanolamine derivatives, methods for their preparation,pharmaceutical compositions containing them and their use as agonists atatypical beta-adrenoceptors (also known as beta-3-adrenoceptors). Suchreceptors have been described for example by J R S Arch et. al., Nature,309, 163-165 (1984); C Wilson et. al., Eur. J. Pharmacol., 100, 309-319(1984); L J Emorine et. al., Science, 245, 1118-1121 (1989); and A.Bianchetti et. al. Br. J. Pharmacol., 100, 831-839 (1990).Phenethanolamine derivatives having activity at atypicalbeta-adrenoceptors are disclosed in, for example, European PatentApplications EP-A-0455006 and EP-A-0543662.

Atypical beta-adrenoceptors belong to the family of adrenoceptors whichmediate the physiological actions of the hormones adrenaline andnoradrenaline. Sub-types of the adrenoceptors, α₁ -, α₂ -, β₁ -, β₂ -and β₃ -(atypical) can be identified on the basis of theirpharmacological properties and physiological effects. Chemical agentswhich stimulate or block these receptors (but not β₃) are widely used inclinical medicine. More recently, emphasis has been placed upon specificreceptor selectivity in order to reduce side effects caused, in part, byinteractions with other receptors.

Atypical beta-adrenoceptors are known to occur in adipose tissue and thegastrointestinal tract.

Atypical beta-adrenoceptor agonists have been found to be particularlyuseful as thermogenic anti-obesity agents and as anti-diabetic agents.Compounds having atypical beta-adrenoceptor agonist activity have alsobeen described as being useful in the treatment of hyperglycaemia, asanimal growth promoters, as blood platelet aggregation inhibitors, aspositive inotropic agents and as antiatherosclerotic agents, and asbeing useful in the treatment of glaucoma.

We have now found a novel class of phenylethanolamine derivatives whichact as agonists at atypical beta-adrenoceptors. GB 9525177.3, which isthe priority document for the present application, describes thesyntheses of the compounds of the invention. WO95/33724, which wasunpublished at the priority date of the present application, describesthe syntheses of compounds which are also of use as agonists at atypicalbeta-adrenoceptors.

The invention therefore provides, in a first aspect, compounds offormula (I): ##STR1## wherein

R¹ represents an aryl group optionally substituted by one or moresubstituents selected from halogen, hydroxy, C₁₋₆ alkoxy, C₁₋₆ alkyl,nitro, cyano, hydroxymethyl and trifluoromethyl;

R² represents hydrogen or C₁₋₆ alkyl;

R³ represents a group A ##STR2## where the ring is substituted by one tofour further substituents selected from C₁₋₆ alkyl, halogen,trifluoromethyl, and C₁₋₆ alkoxy;

or R³ represents a group B ##STR3## where the aromatic ring isoptionally substituted by up to three further substituents selected fromC₁₋₆ alkyl, halogen, trifluoromethyl, and C₁₋₆ alkoxy;

R⁴ represents hydrogen, or C₁₋₆ alkyl;

R⁵ represents ZCH₂ CO₂ H wherein Z represents a bond, or O;

Y represents (CH₂)_(n) where n is 1-3;

and physiologically acceptable derivatives thereof.

Referring to the general formula (I), alkyl includes both straight andbranched chain saturated hydrocarbon groups. Similarly, alkoxy includesboth straight and branched chain groups.

Referring to the general formula (I), aryl includes monocyclic orbicyclic aromatic carbocyclic groups such as phenyl and naphthyl.

Preferably R¹ represents phenyl optionally substituted by one, two orthree substituents selected from halogen, hydroxy, C₁₋₆ alkoxy, C₁₋₆alkyl, nitro, cyano, hydroxymethyl and trifluoromethyl. More preferablyR¹ represents phenyl substituted by a chlorine, fluorine or bromine atomor a methyl or trifluoromethyl group, which atom or group is preferablylocated in the meta position. Most preferably R¹ represents phenylsubstituted by a chlorine atom located in the meta position.

R² is preferably hydrogen or methyl.

Where R³ is group A, preferred substituents are one or more groupsselected from halogen, e.g. fluoro or chloro, methyl, trifluoromethyl,and methoxy.

Where R³ is group B, n is preferably 1 or 2 and the aromatic ring has nofurther substitution.

R⁴ is preferably hydrogen or methyl.

A preferred sub-class of compounds of formula (I) are those where R¹represents phenyl substituted by a chlorine atom located in the metaposition, R² represents hydrogen or methyl, R³ represents a group A andis substituted by one or more groups selected from halogen, methyl,trifluoromethyl, and methoxy, R⁴ represents hydrogen or methyl, R⁵represents CH₂ CO₂ H, or physiologically acceptable derivatives thereof.

It will be appreciated that the above compounds of formula (I) areoptically active. The individual, isolated isomers and mixtures thereof,including racemates, are within the scope of the present invention.Particularly preferred compounds of formula (II) are those wherein theasymmetric carbon atoms in the --CH(OH)-- group and the --CH(CH₃)--group are in the (R)-configuration.

Suitable compounds of formula (I) of the invention are;

(1-{2-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-propyl}-2,3-dihydro-1H-indol-5-yl)-aceticacid;

(1-{2-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-propyl}-1,2,3,4-tetrahydro-quinolin-6-yl)-aceticacid;

(4-{2R-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-propylamino}-2-methyl-phenyl)-aceticacid;

(4-{2R-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-propylamino}-3-methyl-phenyl)-aceticacid;

(4-{2R-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-propylamino}-2-fluoro-phenyl)-aceticacid;

(4-{2R-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-propyiamino}-3-fluoro-phenyl)-aceticacid;

(4-{2R-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-propylamino}-2,3-difluoro-phenyl)-aceticacid;

(5-Chloro-4-{2R-[2-(3-chloro-phenyl)-2R-hydroxy-ethylamino]-propylamino}-2-methoxy-phenyl)-aceticacid;

(4-{2-[2-(3-chloro-phenyl)-2R-hydroxyl-ethyiamino]-ethylamino}-2-methyl-phenyl)-aceticacid;

(4-{2-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-ethylamino}-2,3-difluoro-phenyl)-aceticacid;

(4-{2-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-ethylamino}-2-trifluoromethyl-phenyl)-aceticacid;

(4-{2R-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-propylamino}-2,6-difluoro-phenyl)-aceticacid;

(4-{2-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-ethylamino}-2,3,6-trifluoro-phenyl)-aceticacid;

(4-{2-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-propylamino}-2-trifluoromethyl-phenyl)-aceticacid;

(4-{2R-[2-(3-Chloro-phenyl)-2R-hydroxyl-ethylamino]-propylamino}-2,3,6-trifluoro-phenyl)-aceticacid;

[2-Chloro-4(2-{[2-(3-chloro-phenyl)-2R-hydroxy-ethyl]-amino}-propylamino)-phenyl]-aceticacid;

[5-Chloro-4-(2R-{[2-(3-chloro-phenyl)-2R-hydroxy-ethyl]-amino}-propylamino)-2-methyl-phenyl]-aceticacid;

(4-{2R-[2-(3-Chloro-phenyl)-2R-hydroxy-ethylamino]-propylamino}-2,6-difluoro-phenoxy)-aceticacid;

(4-{2R-[2-(3-Chloro-phenyl)-2R-hydroxy-ethylamino]-propylamino}-3-trifluoromethyl-phenyl)-aceticacid;

or a physiologically acceptable derivative thereof.

Preferred compounds of the invention include:

(4-{2R-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-propylamino}-2,3-difluoro-phenyl)-aceticacid;

or a physiologically acceptable derivative thereof.

By "a physiologically acceptable derivative" is meant anyphysiologically acceptable salt, ester, or salt of such ester, of acompound of formula (I) or any other compound which, upon administrationto the recipient, is capable of providing (directly or indirectly) acompound of formula (I) or an active metabolite or residue thereof.

It will be appreciated by those skilled in the art that the compounds offormula (I) may be modified to provide physiologically acceptablederivatives thereof at any of the functional groups in the compounds offormula (I). Of particular interest as such derivatives are compoundsmodified at the carboxyl function, hydroxyl functions or at aminogroups.

It will be appreciated by those skilled in the art that thephysiologically acceptable derivatives of the compounds of formula (I)may be derivatised at more than one position.

Preferred physiologically acceptable derivatives of the compounds offormula (I) are pharmaceutically acceptable salts thereof.

Pharmaceutically acceptable salts of the compounds of formula (I)include those derived from physiologically acceptable inorganic andorganic acids and bases. Examples of suitable acids includehydrochloric, hydrobromic, sulphuric, nitric, perchloric, fumaric,maleic, phosphoric, glycollic, lactic, salicylic, succinic,toluene-p-sulphonic, tartaric, acetic, citric, methanesulphonic, formic,benzoic, malonic, naphthalene-2-sulphonic and benzenesulphonic acids.Other acids such as oxalic, while not in themselves physiologicallyacceptable may be useful in the preparation of salts useful asintermediates in obtaining compounds of the invention and theirphysiologically acceptable acid addition salts.

Salts derived from appropriate bases include alkali metal (e.g. sodium),alkaline earth metal (e.g. magnesium), ammonium and NR₄ ⁺ (where R isC₁₋₄ alkyl) salts.

The compounds of formula (I) act as agonists at atypicalbeta-adrenoceptors and as such are useful in the treatment of clinicalconditions susceptible to amelioration by administration of an atypicalbeta-adrenoceptor agonist. Such conditions include hyperglycaemia,obesity, hyperlipemia, irritable bowel syndrome and its associated pain,motility dysfunction, excessive gastrointestinal secretion, non-specificdiarrhoea, neurogenic inflammation, regulation of intraocular pressure,triglyceridemia, diabetes, e.g. non-insulin-dependent diabetes mellitus(NIDDM or Type II), such as obese NIDDM and non-obese NIDDM, diabeticcomplications such as retinopathy, nephropathy, neuropathy, cataracts,coronary heart diseases and arteriosclerosis, osteoporosis; andgastrointestinal disorders, particularly inflammatory gastrointestinaldisorders.

Accordingly the present invention provides a method of treatment of amammal, including man, suffering from condition susceptible ofamelioration by an atypical beta-adrenoceptor agonist which methodcomprises administering to the subject an effective amount of a compoundof general formula (I) or a physiologically acceptable derivativethereof.

References in this specification to treatment include prophylactictreatment as well as the alleviation of symptoms.

In a further aspect, the invention provides the use of a compound ofgeneral formula (I) or a physiologically acceptable salt or solvatethereof, for the manufacture of a medicament for the treatment of acondition susceptible of amelioration by an atypical beta-adrenoceptoragonist.

While it is possible that, for use in therapy, a compound of theinvention may be administered as the raw chemical it is preferable topresent the active ingredient as a pharmaceutical formulation.

The invention thus further provides a pharmaceutical formulationcomprising a compound of formula (I) or a physiologically acceptablederivative thereof together with one or more physiologically acceptablecarriers therefor and, optionally, other therapeutic and/or prophylacticingredients. The carrier(s) or excipient(s) must be "acceptable" in thesense of being compatible with the other ingredients of the formulationand not deleterious to the recipient thereof.

Thus the compounds for use according to the present invention may beformulated for oral, buccal, parenteral, rectal or transdermaladministration or in a form suitable for administration by inhalation orinsufflation (either through the mouth or the nose).

For oral administration, the pharmaceutical compositions may take theform of, for example, tablets or capsules prepared by conventional meanswith pharmaceutically acceptable excipients such as binding agents (e.g.pregelatinised maize starch, polyvinylpyrrolidone or hydroxypropylmethylcellulose); fillers (e.g. lactose, microcrystalline cellulose orcalcium hydrogen phosphate); lubricants (e.g. magnesium stearate, talcor silica); disintegrants (e.g. potato starch or sodium starchglycollate); or wetting agents (e.g. sodium lauryl sulphate). Thetablets may be coated by methods well known in the art. Liquidpreparations for oral administration may take the form of, for example,solutions, syrups or suspensions, or they may be presented as a dryproduct for constitution with water or other suitable vehicle beforeuse. Such liquid preparations may be prepared by conventional means withpharmaceutically acceptable additives such as suspending agents (e.g.sorbitol syrup, cellulose derivatives or hydrogenated edible fats);emulsifying agents (e.g. lecithin or acacia); non-aqueous vehicles (e.g.almond oil, oily esters, ethyl alcohol or fractionated vegetable oils);and preservatives (e.g. methyl or propyl-p-hydroxybenzoates or sorbicacid). The preparations may also contain buffer salts, flavouring,colouring and sweetening agents as appropriate.

Preparations for oral administration may be suitably formulated to givecontrolled release of the active compound.

For buccal administration the compositions may take the form of tabletsor lozenges formulated in conventional manner.

The compounds according to the present invention may be formulated forparenteral administration by injection e.g. by bolus injection orcontinuous infusion. Formulations for injection may be presented in unitdosage form e.g. in ampoules or in multi-dose containers, with an addedpreservative. The compositions may take such forms as suspensions,solutions or emulsions in oily or aqueous vehicles, and may containformulatory agents such as suspending, stabilising and/or dispersingagents. Alternatively, the active ingredient may be in powder form forconstitution with a suitable vehicle, e.g. sterile pyrogen-free water,before use.

The compounds according to the present invention may also be formulatedin rectal compositions such as suppositories or retention enemas, e.g.containing conventional suppository bases such as cocoa butter or otherglycerides.

In addition to the formulations described previously, the compounds mayalso be formulated as a depot preparation. Such long acting formulationsmay be administered by implantation (for example subcutaneously,transcutaneously or intramuscularly) or by intramuscular injection.Thus, for example, the compounds according to the present invention maybe formulated with suitable polymeric or hydrophobic materials (forexample as an emulsion in an acceptable oil) or ion exchange resins, oras sparingly soluble derivatives, for example, as a sparingly solublesalt.

A proposed dose of the compounds according to the present invention foradministration to a human (of approximately 70 kg body weight) is 0.1 mgto 1 g, preferably to 1 mg to 100 mg of the active ingredient per unitdose, expressed as the weight of free base. The unit dose may beadministered, for example, 1 to 4 times per day. The dose will depend onthe route of administration. It will be appreciated that it may benecessary to make routine variations to the dosage depending on the ageand weight of the patient as well as the severity of the condition to betreated.

The precise dose and route of administration will ultimately be at thediscretion of the attendant physician or veterinarian.

The compounds of the invention may be prepared by any of the processesknown in the art for the preparation of similar compounds.

For example, according to process (A), compounds of formula (I) may beprepared by reaction of a compound of formula (Ia) ##STR4## wherein R¹,R², and, are as defined as for formula (I), R³ represents R³ where theacidic group is protected by an alkyl ester, and R^(a) and R^(b) areprotecting groups, by deprotection of the protecting groups in asuitable mixture such as 6M hydrochloric acid in tetrahydrofuran.

According to process (B), compounds of formula (I) may be prepared byreaction of a compound of formula (II) with a compound of formula (III):##STR5## wherein R¹, R², R³, R^(a), and R^(b) are as defined above, inthe presence of a reducing agent, followed by removal of the protectinggroups.

Compounds of formula (II) may be prepared by reaction of compounds offormula (IV), with an amine acid salt of formula (V). ##STR6## whereinR¹, R², and R^(b) are as defined herein before, and R^(c) is a suitablealkyl group for protection, in the presence of a reducing agent.Following protection of the nitrogen, the ester is reduced by a suitablereducing agent such as di-isobutyl aluminium hydride.

Where R³ is group A, compounds of formula (III) may be prepared fromcompounds of formula (VI) ##STR7## where the aromatic ring is optionallysubstituted as defined for R³, by treatment with a mixed ester, e.g.methyl, benzyl, of malonic acid at elevated temperature in a suitablesolvent such as N,N dimethylformamide, followed by treatment with asuitable reducing agent.

Suitable reducing agents of use in the reactions include hydrogen in thepresence of a catalyst, such as a noble metal catalyst, for examplepalladium, platinum or platinum oxide, Raney-nickel or hydride reducingagents such as borohydrides, for example sodium borohydride sodiumtriacetoxyborohydride or sodium cyanoborohydride. Suitable reactionconditions will be readily apparent to those skilled in the art and arefurther illustrated by the accompanying examples.

Compounds of formula (III) where R³ represents group B, (IV), (V) and(VI) are known compounds or may be prepared from known compounds bystandard procedures well known to those skilled in the art.

The protecting groups used in the preparation of compounds of formula(I) may be used in conventional manner. See for example `ProtectiveGroups in Organic Chemistry` Ed. J. F. W. McOmie (Plenum Press 1973) or`Protective Groups in Organic Synthesis` by Theodora W Greene and P M GWuts (John Wiley and Sons 1991).

Conventional amino protecting groups may include for example aralkylgroups, such as benzyl, diphenylmethyl or triphenylmethyl groups; andacyl groups such as N-benzyloxycarbonyl or t-butoxycarbonyl.

Conventional oxygen protecting groups may include for example alky silylgroups, such as trimethylsilyl, or tert-butyldimethylsilyl; alkyletherssuch as tetrahydropyranyl, or tert-butyl; or esters such as acetate.

Removal of any protecting groups present may be achieved by conventionalprocedures.

Atypical beta-adrenoceptor agonists are compounds which demonstrate apharmacological response mediated at atypical beta-adrenoceptors. Thisactivity has been be measured as the ability to stimulate lipolysis byrat adipocytes at sub-micromolar concentrations, in a response that isresistant to blockade by standard beta-adrenoceptor blocking drugs suchas propranolol.

A particularly useful method for determining agoinst activity at humanatypical beta-adrenoceptors involves the use of Chinese hamster ovarian(CHO) cells transfected with the human beta-3-adrenoceptor according toMethod 2. The cell lines may also be transfected with human beta-1- andbeta-2- adrenoceptor in a similar manner to provide a method ofdetermining the selectivity of the compounds of the invention at thethree receptors.

METHOD 1 Cell Culture

General cell culture guidelines are observed (Fershney, R. A. (1987)Culture of animal cells: A manual of basic technique. Wiley-Liss, Inc.,N.Y.). A standard cell culture incubator is used (37° C., 5% CO₂ in air,95% relative humidity). H β₃ CHO cells are grown in 75 ml flasks in MEMαmedium containing 9% FCS & 125 μg/ml G418. One confluent flask of cellsis trypsinised and resuspended in 80 ml of culture medium; 1 ml of thecell suspension is added to each well of three 24-well plates. Theplates are then incubated for 1 day.

Experimental method:

The medium is aspirated from each well, and the well rinsed withphosphate-buffered saline (PBS, this is then aspirated). 1 ml of MEMα(no FCS or G418, 300μ M IBMX) is added to each well. Antagonists, ifrequired, are added at this stage. The plate is then placed back in theincubator for 30 min. Drugs are then added to the wells (10 μl, 100xrequired final concentration), the plate gently swirled to mix thedrugs, and the plate placed back in the incubator for 30 min. The mediumis then aspirated from each well, the well rinsed with PBS, and 0.5 mlperchloric acid (6% v/v in distilled water, 2-5° C.). The plate is lefton ice for 30 min. The perchloric acid (containing cAMP) is transferredto a clean 24-well plate and the acid neutralised by addition ofsaturated KHCO₃ solution (200 μl) to each well. The plate is thenswirled and frozen (-20° C.) until cAMP is assayed. cAMP is assayedusing an enzyme-immunoassay kit (Amersham).

The relative potency of each test agonist (EPMR) is compared toisoprenaline as follows: ##EQU1## wherein EC₅₀ is the molarconcentration of agonist which produces 50% of the maximum possibleresponse for that agonist.

Using the non-selective beta-adrenoceptor agonist isoprenaline as areference agonist, compounds selective for atypical beta-adrenoceptorsshould preferably be a minimum of 10-30 times less potent thanisoprenaline at β₁ - or β₂ -adrenoceptors and, more preferably, 300-1000times less potent than isoprenaline at β₁ - or β₂ -adrenoceptors.

An experimental model in which atypical beta-adrenoceptor agonists maybe shown to be of use in the treatment of gastrointestinal disorders isdescribed below as Method 3. The procedure is based upon that describedby H. Satoh et. al., Gastroenterology, 81, 719-725 (1981) in which theeffect of compounds on indomethacin-induced gastric antral lesions inthe re-fed rat is investigated. Indomethacin is an example of the classof compound known as non-steroidal anti-inflammatory drugs (NSAIDs), theuse of which is frequently associated with gastrointestinal ulcers.

METHOD 2

Food (but not water) is withheld from female random hooded rats (70-120g) for 24 hours and then the rats are re-fed with Rat and Mouse No. 1Maintenance Diet. After 1 hour of access to food, the rats are dosedorally with either the test compound or solvent (0.5% w/v methylcellulose in water). 30 minutes later, indomethacin (60 mg/kg; dissolvedin 1% w/v NaHCO₃ in saline) is administered as a single subcutaneousinjection at the back of the neck. Subsequently, the rats are allowedfood, but water is withheld, and the animals are humanely killed bycervical dislocation at 6 hours post dose. Control animals received asingle subcutaneous dose of the appropriate solvent.

The rat's stomach is removed (with a small amount of duodenum attached),opened along the greater curvature and the contents removed by washingwith 0.9% w/v sodium chloride solution (saline). The opened stomach ispinned out (mucosal surface uppermost) on a polystyrene mat and the areaof damage assessed by placing a grid (composed of 1 mm squares) over theantral region. Antral damage appears as discrete black or dark brownulcers. The total area of antral damage is then expressed as apercentage of the total surface area of the antrum.

The protective effect of the test compound on indomethacin-inducedantral damage is calculated as a percentage using the followingequation: ##EQU2##

The invention is further illustrated by the following intermediates andexamples. All temperatures are in degrees centigrade.

INTERMEDIATE 1

(R)-(3-chloro-phenyl)-hydroxy-acetic acid methyl ester

A solution of (R)-(3-chloro-phenyl)-hydroxy-acetic acid (19.98 g) inmethanol (250 ml) containing concentrated sulphuric acid (1 ml) washeated under reflux for 6.5 h. The solution was cooled, neutralised withaqueous sodium bicarbonate solution, and concentrated. The residue wasdissolved in ethyl acetate, washed with aqueous sodium bicarbonatesolution, dried, and evaporated to give the title compound (21.13 g) asa pale-yellow oil.

[a]_(D) -104° (c 1.00 MeOH)

INTERMEDIATE 2

(R)-(3-chloro-phenyl)-(tert-butyl-dimethyl-silanoxy)-acetic acid methylester

A solution of (R)-(3-chloro-phenyl)-hydroxy-acetic acid methyl ester(21.0 g), imidazole (14.25 g), and tert-butyidimethylsilyl chloride(25.0 g) in N,N-dimethylformamide (250 ml) was stirred at roomtemperature for 18 h. The mixture was poured into water (2.5 l) andextracted with ethyl acetate. The combined extracts were washed withwater and saturated brine, dried, and concentrated. The residue waspurified by flash chromatography, eluting with cyclohexane:ethyl acetate(9:1) to give the title compound as a colourless oil (32.63 g).

[a]_(D) -55.4° (c 1.21 MeOH)

INTERMEDIATE 3

(R)-(3-chloro-phenyl)-(tert-butyl-dimethyl-silanoxy)-acetaldehyde

To a stirred solution of(R)-(3-chloro-phenyl)-(tert-butyl-dimethyl-silanoxy)-acetic acid methylester (4.0 g) in anhydrous ether (10 ml) and maintained at <-65° wasadded dropwise a 1.5M solution of di-isobututylaluminium hydride intoluene (10 ml). When addition was complete the solution was stirred at-65° for a further hour, then quenched with methanol (10 ml). Themixture was allowed to attain room temperature when silica (20 g) wasadded. Solvent was removed under reduced pressure, and the residue waspurified by flash chromatography, eluting with cyclohexane:ethyl acetate(9:1) to give the title compound as a colourless liquid (3.1 g).

[a]_(D) -45.3° (c 1.50 MeOH)

INTERMEDIATE 4

2R-[2R-(tert-Butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethylamino]-propionicacid methyl ester

(R)-(3-chloro-phenyl)-(tert-butyl-dimethyl-silanoxy)-acetaldehyde (2.32g) was added to a stirred solution of (R)-2-aminopropionic acid methylester hydrochloride (1.13 g) in dichioromethane (50 ml). The solutionwas stirred for 15 min, then sodium triactetoxyborohydride (3.45 g) wasadded, and the mixture was stirred a further 18 h. The solution waswashed with aqueous sodium bicarbonate solution, then the organic phasewas dried and concentrated. The residue was purified by chromatography,eluting with cyclohexane:ethyl acetate (9:1) to give the title compoundas a colourless oil (2.42 g)

[a]_(D) -29.4° (c 1.36 MeOH)

Similarly prepared was:

INTERMEDIATE 5

[2R-(tert-Butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethylamino]-aceticacid methyl ester as a colourless oil (1.26 g),

Assay: Found: C 56.9; H 7.95; N 3.9% C₁₇ H₂₈ ClNO₃ Si requires C 57.1; H7.8; N 3.9% from amino-acetic acid methyl ester hydrochloride (1.05 g)and (R)-(3-chloro-phenyl)-(tert-butyl-dimethyl-silanoxy)-acetaldehyde(2.16 g).

INTERMEDIATE 6

{2R-(tert-Butoxycarbonyl)-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propionicacid methyl ester

A mixture of2R-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethylamino]-propionicacid methyl ester (2.15 g) and di-t-butyl pyrocarbonate (1.37 g) washeated at 80-100° for 1 h. The mixture was cooled and purified bychromatography eluting with cyclohexane:ethyl acetate (19:1) to give thetitle compound as a colourless oil (2.70 g).

Assay: Found: C 58.4; H 8.1; N 3.0% C₂₃ H₃₈ ClNO₅ Si requires C 58.5; H8.1; N 3.0%

Similarly prepared was:

INTERMEDIATE 7

{(tert-Butoxycarbonyl)-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-aceticacid methyl ester as a colourless oil (1.55 g) from[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethylamino]-aceticacid methyl ester (1.20 g)

[a]_(D) -25.2° (c 1.3 MeOH)

INTERMEDIATE 8

{2R-(tert-Butoxycarbonyl)-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propionaldehyde

1.5M di-isobutylaluminium hydride in toluene (8.1 ml) was added dropwiseto a stirred, cooled solution of the{2R-(tert-butoxycarbonyl)-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propionicacid methyl ester (2.30 g) in toluene (50 ml) at such a rate that thereaction temperature did not rise above -70°. The solution was stirred 1h at this temperature, then quenched with methanol (10 ml). The mixturewas preabsorbed on silica and purified by chromatography eluting withcyclohexane:ethyl acetate (9:1) to give the title compound as acolourless oil (1.45 g).

C₂₂ H₃₆ ClNO₄ Si: MH⁺ 443

Similarly prepared was:

INTERMEDIATE 9

{(tert-Butoxycarbonyl)-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-acetaldehydeas a colourless gum (0.37 g),

n.m.r. (CDCl₃): d -0.15 (d, 3H), 0.05 (d, 3H), 0.90 (s, 9H), 1.45 (d,9H), 2.9-3.2 (m, 1H), 3.4-3.65 (m, 1H), 3.70 (s, 3H), 3.75-4.15 (m, 2H),4.8-5.0 (m, 1H), 7.05-7.35 (m, 4H).

from{(tert-butoxycarbonyl)-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chlorophenyl)-ethyl]-amino}-aceticacid methyl ester (0.50 g).

INTERMEDIATE 10

(2,3-Dihydro-1H-indol-5-yl)-acetic acid methyl ester

A suspension of (1-acetyl-2,3-dihydro-1H-indol-5-yl)-acetic acid methylester (0.60 g) in 2M hydrochloric acid (15 ml) was heated under refluxfor 5 h. The mixture was evaporated to dryness. The residue wasdissolved in methanol (20 ml) and treated with concentrated sulphuricacid (5 drops). The solution was stirred at room temperature for onehour, then concentrated. The residue was partitioned between ethylacetate and sodium carbonate solution. The organic phase was dried andevaporated and the residue purified by chromatography eluting withcyclohexane:ethyl acetate (2:1) to give the title compound as a brownoil (0.40 g).

Assay Found: C 68.9; H 6.6; N 7.2% C₁₁ H₁₃ NO₂ requires C 69.1; H 6.8; N7.3%

INTERMEDIATE 11

(4-Amino-2-methyl-phenyl)-acetic acid methyl ester

Sodium hydride (1.29 g of 60% dispersion in oil) was added portionwiseto a stirred solution of malonic acid, methyl benzyl ester (5.84 ml) indry N,N-dimethylformamide (40 ml). After 2.5 h 2-fluoro-5-nitro-toluene(5.0 g) was added, and the mixture was warmed to 100° for 18 h. Themixture was cooled then partioned between ethyl acetate and water. Theaqueous phase was separated and extracted further with ethyl acetate.The combined organic extracts were dried, concentrated, and purified bychromatography eluting with cyclohexane:ethyl acetate (6:1) to give2-(2-methyl-4-nitro-phenyl)-malonic acid benzyl ester methyl ester,admixed with the starting malonate ester, as a yellow oil (4.60 g).

The crude mixture (4.2 g), ammonium formate (7.1 g), and 10% palladiumon carbon (0.45 g) in methanol (150 ml) was heated under reflux for 2 hunder an atmosphere of nitrogen. The mixture was cooled, filtered, andthe filtrate was concentrated. Chromatography of the residue elutingwith cyclohexane:ethyl acetate (2:1) gave the title compound (1.55 g) asa yellow oil.

Assay Found: C 66.55; H 7.6; N 7.6% C₁₀ H₁₃ NO₂ requires C 67.0; H 7.3;N 7.8%

n.m.r. (CDCl₃): d values include 2.22 (s, 3H), 3.55 (s, 2H), 3.59 (broads, 2H), 3.68 (s, 3H), 6.51 (s+d, 2H), 6.98 (d, 1H)

Similarly prepared were:

INTERMEDIATE 12

(4-Amino-3-methyl-phenyl)-acetic acid methyl ester as a pale brown oil(0.901 g),

n.m.r. (CDCl₃): d values include 2.12 (s, 3H), 3.49 (s, 2H), 3.58 (broads, 2H), 3.66 (s, 3H), 6.60 (d, 1H), 6.92 (s+d, 2H).

from 5-fluoro-2-nitrotoluene (2.5 g) and malonic acid, benzyl estermethyl ester (5.84 ml)

INTERMEDIATE 13

(4-Amino-2-fluoro-phenyl)-acetic acid methyl ester as a pale brown oil(1.43 g)

n.m.r. (CDCl₃): d values include 3.57 (s, 2H), 3.74 (broad s, 2H), 3.69(s, 3H), 6.32-6.49 (m, 2H), 7.00 (t, 1H).

from 3,4-difluoro-nitrobenzene (3.48 ml) and malonic acid, benzyl estermethyl ester (5.69 ml)

INTERMEDIATE 14

(4-Amino-3-fluoro-phenyl)-acetic acid methyl ester as a yellow oil (0.34g)

n.m.r. (CDCl₃): d values include 3.50 (s, 2H), 3.69 (s, 3H), 6.66-6.90(m, 3H).

from 2,4-difluoro-nitrobenzene (3.33 ml) and malonic acid, benzyl estermethyl ester (6.0 ml)

INTERMEDIATE 15

(4-Amino-2,3-difluoro-phenyl)-acetic acid methyl ester as a pale yellowoil (1.08g),

n.m.r. (CDCl₃): d values include 3.59 (s, 2H), 3.71 (s, 3H), 3.80 (broads, 2H), 6.50 (t, 1H), 6.78 (t, 1H)

from 2,3,4-trifluoro-nitrobenzene (3.59 ml) and malonic acid, benzylester methyl ester (5.69 ml)

INTERMEDIATE 16

(4-Amino-2,3,6-trifluoro-phenyl)-acetic acid methyl ester as colourlesscrystals (1.35 g)

n.m.r. (CDCl₃): d values include 3.72 (s, 3H), 3.60 (s, 2H), 6.30 (m,1H).

from 2,3,4,6-tetrafluoro-nitrobenzene (3.23 g) and malonic acid, benzylester methyl ester (3.63 g).

INTERMEDIATE 17

(4-Amino-2-bromo-phenyl)-acetic acid methyl ester as a yellow oil (0.104g)

n.m.r. (CDCl₃): d values include 3.69 (s. 2H), 3.71 (s, 3H), 6.59 (dd,1H), 6.86 (m, 1H), 7.06 (d, 1H),

from 3-bromo-4-fluoro-nitrobenzene (6.91 g) and malonic acid, benzylester methyl ester (5.69 ml)

INTERMEDIATE 18

(4-Amino-2-trifluoromethyl-phenyl)-acetic acid methyl ester

A solution of (4-amino-2-trifluoromethyl-phenyl)-acetic acid (0.082 g)in methanol (10 ml) containing concentrated sulphuric acid (0.1 ml) washeated under reflux for 2 h. The solution was cooled, concentrated, andpartitioned between ethyl acetate and aqueous sodium carbonate solution.The organic phase was separated, dried, and concentrated to give thetitle compound as a yellow solid (0.089 g)

C₁₀ H₁₀ F₃ NO₂ : MH⁺ 234

INTERMEDIATE 19

[1-(2-{tert-Butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanoxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propyl)-2,3-dihydro-1H-indol-5-yl]-aceticacid methyl ester

A solution of (2,3-dihydro-1H-indol-5-yl)-acetic acid methyl ester (0.10g) and{2R-(tert-butoxycarbonyl)-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propionaldehyde(0.20 g) in dichloromethane (15 ml) containing acetic acid (0.031 ml)was stirred at room temperature for 40 min, then the reaction mixturewas cooled to 0°, and sodium triacetoxyborohydride (0.19 g) was added.The mixture was stirred at room temperature for two days, then washedwith aqueous sodium bicarbonate solution. The organic phase was dried,concentrated, and the residue purified by chromatography eluting withcyclohexane:ethyl acetate (9:1) to give the title compound as a yellowgum (0.22 g)

Assay Found: C 64.1; H 7.6; N 4.4% C₃₃ H₄₉ ClN₂ O₅ requires C 64.2; H7.95; N 4.5%

Chiral HPLC and n.m.r. confirm the compound to be a mixture of RR and RSisomers (1:1).

Similarly prepared were:

INTERMEDIATE 20

[1-(2{-tert-Butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanoxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propyl)-1,2,3,4-tetrahydro-quinolin-6-yl]-aceticacid methyl ester as a yellow oil (0.51 g)

n.m.r. (CDCl₃): d values include 0.88 (s, 9H), 3.46 (s, 2H), 3.67 (s,3H), 6.84 (broad s, 1H), 6.91 (d, 2H), C₃₄ H₅₁ ClN₂ O₅ Si: MH⁺ 631

from (1,2,3,4-tetrahydro-quinolin-6-yl)-acetic acid methyl ester (0.20g) and{2R-(tert-butoxycarbonyl)-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propionaldehyde(0.55 g).

INTERMEDIATE 21

[4-(2-{tert-Butoxycarbonyl-[2-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propylamino)-2-methyl-phenyl]-aceticacid methyl ester as a pale yellow oil (0.22 g),

Assay Found: C 63.2; H 8.6; N 4.7% C₃₂ H₄₉ ClN₂ O₅ Si requires C 63.5; H8.2; N 4.6%

from 4-amino-2-methyl-phenylacetic acid methyl ester (0.11 g) and{2R-(tert-butoxycarbonyl)-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propionaldehyde(0.25 g).

INTERMEDIATE 22

[4-(2R-{tert-Butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propylamino)-3-methyl-phenyl]-aceticacid methyl ester as a pale yellow gum (0.25 g),

Assay Found: C 63.4; H 8.2; N 4.45% C₃₂ H₄₉ ClN₂ O₅ Si requires C 63.5;H 8.2; N 4.6%

from 4-amino-3-methylphenyl-acetic acid methyl ester (0.10 g) and{2R-(tert-butoxycarbonyl)-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propionaldehyde(0.25 g)

INTERMEDIATE 23

[4-(2R-{tert-Butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propylamino)-2-fluoro-phenyl]-aceticacid methyl ester as a colourless oil (0.25 g),

Assay Found: C 61.1; H 7.8; N 4.4% C₃₁ H₄₆ ClFN₂ O₅ Si requires C 61.1;H 7.6; N 4.6%

from 4-amino-2-fluoro-phenylacetic acid methyl ester (0.10 g) and{2R-(tert-butoxycarbonyl)-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propionaldehyde(0.25 g)

INTERMEDIATE 24

[4-(2R-{tert-Butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propylamino)-3-fluoro-phenyl]-aceticacid methyl ester as a pale yellow gum (0.56 g),

Assay Found: C 61.2; H 7.8; N 4.7% C₃₁ H₄₆ ClFN₂ O₅ Si requires C 61.1;H 7.6; N 4.6%

from 4-amino-3-fluoro-phenylacetic acid methyl ester (0.27 g) and{2R-(tert-butoxycarbonyl)-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propionaldehyde(0.66 g)

INTERMEDIATE 25

[4-[2R-{tert-Butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propylamino)-2,3-difluoro-phenyl]-aceticacid methyl ester as a colourless oil (0.50 g),

Assay Found: C 59.9; H 7.4; N 4.4% C₃₁ H₄₅ ClF₂ N₂ O₅ Si requires C59.4; H 7.2; N 4.5%

from 4-amino-2,3-difluoro-phenylacetic acid methyl ester (0.3 g) and{2R-(tert-butoxycarbonyl)-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propionaldehyde(0.50 g).

INTERMEDIATE 26

[4-(2R-{tert-Butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propylamino)-5-chloro-2-methoxy-phenyl]-aceticacid methyl ester as a colourless oil (0.04 g),

C₃₂ H₆₈ Cl₂ N₂ O₆ Si: MH⁺ 656

from 4-amino-5-chloro-2-methoxy-phenylacetic acid, methyl ester (0.05 g)and{2R-(tert-butoxycarbonyl)-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propionaldehyde(0.1 g).

INTERMEDIATE 27

[4-(2-{tert-Butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-ethylamino)-2-methyl-phenyl]-aceticacid methyl ester as a yellow gum

C₃₁ H₄₇ ClN₂ O₅ : MH⁺ 591

from (4-amino-2-methyl-phenyl)-acetic acid methyl ester (0.20 g) and{2R-(tert-butoxycarbonyl)-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-acetaldehyde(0.525 g)

INTERMEDIATE 28

[4-(2-{tert-Butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-ethylamino)-2-trifluoromethyl-phenyl]-aceticacid methyl ester as a colourless gum

C₃₁ H₄₄ F₃ ClN₂ O₅ : MH⁺ 645

from (4-amino-2-trifluoromethyl-phenyl)-acetic acid methyl ester (0.087g) and{2R-(tert-butoxycarbonyl)-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-acetaldehyde(0.224 g).

INTERMEDIATE 29

[4-(2R-{tert-Butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propylamino)-2-trifiuoromethyl-phenyl]-aceticacid methyl ester as a colourless gum (0.307 g)

C₃₁ H₄₄ F₃ ClN₂ O₅ : MH⁺ 659

from (4-amino-2-trifluoromethyl-phenyl)-acetic acid methyl ester (0.150g) and{2R-(tert-butoxycarbonyl)-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-acetaldehyde(0.313 g).

INTERMEDIATE 30

[4-(2-{tert-Butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-ethylamino)-2,3-difluoro-phenyl]-aceticacid methyl ester as a colourless gum (0.12 g)

n.m.r. (CDCl₃): d values include -0.11 (s, 3H), 0.03 (3, 3H), 0.88 (s,9H), 3.56 (s, 2H), 3.70 (s, 3H), 6.35 (t, 1H), 6.81 (t, 1 H), 7.37(broad s, 1H).

from (4-amino-2,3-difluoro-phenyl)-acetic acid methyl ester (0.10 g) and{2R-(tert-butoxycarbonyl)-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-acetaldehyde(0.30 g).

INTERMEDIATE 31

[4-(2R-{tert-Butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propylamino)-2,3,6-trifluoro-phenyl]-aceticacid methyl ester as a colourless gum (0.293 g),

n.m.r. (CDCl₃): d values include -0.11 (s, 3H), 0.032 (s, 3H), 0.88 (s,9H), 3.02-3.29 (m, 4H), 3.59 (s, 2H), 3.71 (s, 3H), 6.17 (m, 1H),

from{2R-(tert-Butoxycarbonyl)-[2R-(tert-Butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propionaldehyde(0.83g) and (4-Amino-2,3,6-trifluoro-phenyl)-acetic acid methyl ester(0.33 g)

INTERMEDIATE 32

[4-(2R-{tert-Butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propylamino)-2-chloro-phenyl]-aceticacid methyl ester as a yellow oil (400 mg), from{2R-(tert-butoxycarbonyl)-[2R-(tert-butyl-dimethyl-silanoxy)-2-(3-chloro-phenyl)-ethyl]-amino}propionaldehyde(0.36 g, 0.8 mmol) and (4-amino-2chloro-phenyl)acetic acid methyl ester(0.23 g, 1.19 mmol)

INTERMEDIATE 33

[4-(2R-{tert-Butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propylamino)-5-chloro-2-methyl-phenyl]-aceticacid methyl ester as a colorless oil (230 mg)

from{2R-(tert-butoxycarbonyl)-[2R-(tert-butyl-dimethyl-silanoxy)-2-(3-chloro-phenyl)-ethyl]-amino}propionaldehyde(0.35 g) and (4-amino-2-methyl-3-chloro-phenyl)acetic acid methyl ester(0.26 g).

INTERMEDIATE 34

(4-{2R-{tert-Butoxycarbonyl-[2R-(tertbutyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propylamino)-2,6-difluoro-phenyoxy)-aceticacid methyl ester(454 mg),

TLC (2:1 hexanes: ethyl acetate) Rf 0.6

from (4-amino-2,6-difluoro-phenoxy)-acetic acid methyl ester (314 mg)and{2R-(tert-butoxycarbonyl)-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propionaldehyde(380 mg)

INTERMEDIATE 35

2-(4-nitro-2chlorophenyl)-malonic acid dimethyl ester

3-Chloro-4-fluoronitobenzene (1.9 g) and dimethylmalonate (1.26 mL) weredissolved in N-methylpyrrolidinone (50 mL). Sodium hydroxide (0.92 g)was added and the solution heated at 80° for 2 h. The reaction wascooled and 1 N hydrochloric acid added. The mixture was extracted withethyl acetate, concentrated and the residue purified by chromatographyon silica gel eluting with hexane/ethyl acetate (9:1) to give the titlecompound (2.45 g)

INTERMEDIATE 36

2-(4 nitro-2-chlorophenyl)acetic acid methyl ester

2-(4-nitro-2chlorophenyl)-malonic acid dimethyl ester was dissolved inDMSO, then lithium chloride (0.759 g), and water (0.15 mL) were added.The mixture was heated to 100° C. for 3h. The reaction was cooled andthe product purified by chromatography on silica gel eluting withhexane/ethyl acetate (9:1) to give the title compound as a yellow oil(1.77 g).

INTERMEDIATE 37

(4-Amino-2-chloro-phenyl)-acetic acid methyl ester

2-(4 nitro-2-chlorophenyl )acetic acid methyl ester was reduced with tinchloride in methanol at reflux for 2 h. The solution was poured over iceand neutralized with saturated sodium bicarbonate. The mixture wasextracted with methylene chloride, dried (MgSO4), and concentrated. Theoil was purified by silica gel chromatography eluted with hexane/ethylacetate(9:1 to 8:2) to yield the title compound (0.81 g).

1H NMR (400 mHz, CDCl₃ ) d 7.0 (d, 1H), 6.7 (s, 1H), 6.5 (d, 1H), 3.74(s, 2H), 3.68 (s, 3H), 3.63 (s, 2H)

INTERMEDIATE 38

2-(4-nitro-5-chloro-2-methyl)phenyl acetic acid methyl ester

4-Chloro-2-fluoro-5-nitrotoluene (1.0 g) and dimethylmalonate (0.61 mL)were dissolved in N-methylpyrrolidinone (40 mL). Sodium hydroxide (0.45g) was added and the solution heated at 80° for 4 h. Reaction was cooledand 1 N hydrochloric acid added. The mixture was extracted with ethylacetate and concentrated to give 1.52 g of crude product (96%). Theabove product was dissolved in DMSO and lithium chloride (0.45 g), andwater (0.90 mL) were added. The mixture was heated to 100° C. for 4 h.and allowed to stir at room temperature for 18 h. Hydrochloric acid (1N) was added and the solution extracted with ethyl acetate, dried,filtered, concentrated and the residue purified by chromatography onsilica gel to give the title compound as an oil (0.53 g)

INTERMEDIATE 39

(4-Amino-2-chloro-5-methyl-phenyl)-acetic acid methyl ester

(4-nitro-2-chloro-5-methyl)phenyl acetic acid methyl ester was reducedwith tin chloride in ethanol at 70° C. for 1 h. The solution was cooledand poured over ice and neutralized with saturated sodium bicarbonate.The mixture was extracted with ethyl acetate, dried, and concentrated.The resulting oil was purified by silica gel chromatography eluting withhexane/ethyl acetate(9:1) to yield the title compound (0.446 g).Analysis by NMR indicated a mixture of methyl and ethyl esters.

¹ H NMR (400 mHz, CDCl₃) d 7.09 (s, 1H), 6.57 (s, 1H), 4.1 (q), 3.66 (s,2H), 3.46 (d), 2.17 (s, 3H), 1.2 (t)

INTERMEDIATE 40

2-(4-amino-3-trifluoromethyl-phenyl]-malonic acid methyl ester

To a solution of benzylmethyl malonate (0.99 g) in anhydrousN,N-dimethylformamide (50 mL) was added 60% NaH in mineral oil (0.18 g)in portions. After 20 min, 4-fluoro-2-trifluoromethylnitrobenzene (1.0g) in N,N-dimethylformamide (20 mL) was added dropwise via additionfunnel. Following the addition, the mixture was heated at 90° C. for 3h. The mixture was allowed to cool to ambient temperature, and dilutedwith water. The mixture was extracted with ethyl acetate and thecombined organic layers were dried over Na₂ SO₄, filtered andconcentrated to give the intermediate benzyl methyl ester as an orangeoil. This material was stirred under 1 atmosphere hydrogen gas in 30 mLmethanol with 0.20 b 10% Pd/C catalyst for 4.5 h. Filtration through apad of celite and concentration of the filtrate afforded the titlecompound (340 mg) as a red oil.

C₁₀ H₁₀ F₃ N₁ O₂ : M-H 232

INTERMEDIATE 41

[4-(2R-Amino-propylamino)-3-trifluoromethyl-phenyl]-acetic acid methylester

A mixture of 2-(4-amino-3-trifluoromethyl-phenyl)-malonic acid methylester (356 mg), (2R)-(-)-tert-butyoxycarbonyl-amino-1-propionaldehyde(341 mg) and acetic acid (3 drops) in anhydrous dichloromethane (20 mL)was stirred for 10 min. Sodium triacetoxyborohydride (780 mg) was added,and the mixture was stirred at ambient temperature for 24 h. The mixturewas diluted with dichloromethane and washed with saturated aqueoussodium bicarbonate. The organic layer was dried over anhydrous Na₂ SO₄,and concentrated. The residue was purified by chromatography elutingwith hexane: ethyl acetate (5:1) to afford a pale yellow oil.Concentration of the relevant fractions gave a material which wasdissolved in dichloromethane (40 mL) and trifluoroacetic acid (4 mL).The mixture was stirred at ambient temperature for 3 h, andconcentrated. The residue was partitioned between aqueous sodium acetateand ethyl acetate. The ethyl acetate layer was concentrated to afford aresidue which was purified by silica gel chromatography eluting withethyl acetate: methanol (5:1) to afford, after concentration of therelevant fractions, the title compound (242 mg) as a yellow oil.

C₁₃ H₁₇ N₂ O₂ F₃ : MH⁺ 291

INTERMEDIATE 42

(4-{2R-[2-(3-Chloro-phenyl)-2R-hydroxy-ethylamino]-propylamino}-3-trifluoromethyl-phenyl)-aceticacid methyl ester

A mixture of [4-(2R-amino-propylamino)-3-trifluoromethyl-phenyl]-aceticacid methyl ester (255 mg) and (R)-3-chlorostyrene oxide (90 mg) was innitromethane (4 mL) was heated at 105° C. for 36 h. The mixture wasallowed to cool to ambient temperature, and concentrated to afford aresidue which was purified by silica gel chromatography eluting withhexane: ethyl acetate (2:1) followed by 1:1 hexanes: ethyl acetate toafford after concentration of the relevant fractions the title compound(75.1 mg) as a tan oil.

C₂₁ H₂₄ Cl F₃ N₂ O₃ : MH⁺ 445

INTERMEDIATE 43

(4-Nitro-2,6-difluoro-phenoxy)-acetic acid methyl ester

A mixture of 4-nitro-2,6-difluoro-phenol (5.08 g, 29.0 mmol) and cesiumcarbonate (10.3 g, 31.7 mmol) in 100 mL acetonitrite was treated withmethyl bromoacetate (3.0 mL, 31.7 mmol). The mixture was heated atreflux for 2 h. The mixture was allowed to cool to ambient temperature,and partitioned between water and ethyl acetate. The organic layer wasseparated and dried over sodium sulfate, filtered, and concentrated toafford material which was purified by silica gel chromatography elutingwith hexane: ethyl acetate (2:1). Concentration of the relevantfractions afforded the title compound (6.11 g) as a white solid.

n.m.r (CDCl₃): d 3.83 (s, 3H), 4.98 (s, 2H), 7.85-7.98 (m, 2H).

INTERMEDIATE 44

(4-Amino-2,6-difluoro-phenoxy)-acetic acid methyl ester

A slurry of (4-nitro-2,6-difluoro-phenoxy)-acetic acid methyl ester (737mg, 2.98 mmol) and 10% palladium on carbon (200 mg) in tetrahydrofuran(40 mL) was stirred under 1 atmosphere of hydrogen gas for 7 h. Themixture was flushed with nitrogen, and filtered through a pad of celiteto afford the title compound as a pale tan oil (646 mg) that solidifiedon standing.

n.m.r (CDCl₃): d 3.68 (bs, 2H), 3.78 (s, 3H), 4.59 (s, 2H), 6.17-6.21(m, 2H).

EXAMPLE 1

(1-{2-[2-(3-Chloro-phenyl)-2R-hydroxyl-ethylamino]-propyl}-2,3-dihydro-1H-indol-5-yl)-aceticacid

[1-(2-{tert-Butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanoxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propyl-2,3-dihydro-1H-indol-5-yl]-aceticacid methyl ester (0.20 g) was dissolved in tetrahydrofuran (5 ml) andthe solution was treated with 6N hydrochloric acid (5 ml) for 72 h. Thesolution was evaporated to dryness and the residue purified bychromatography with Sorbsil C60 eluting with thechloroform:methanol:0.880 ammonium hydroxide (10:5:1) to give the titlecompound as a yellow solid (0.08 g).

Assay Found: C 61.0; H 6.5; N 6.8% C₂₁ H₂₅ ClN₂ O₃.1.25H₂ O requires C61.3; H 6.7; N 6.8%

n.m.r. (DMSO-d₆): d values include 1.05 (m, 3H), 2.61-3.30 (m, 5H), 4.70(m, 1H), 6.41 (m, 1H), 6.85-6.95 (m, 2H), 7.25-7.45 (m, 4H).

Similarly prepared were:

EXAMPLE 2

(1-{2-[2-(3-Chloro-phenyl)-2R-hydroxyl-ethylamino]-propyl}-12,3,4-tetrahydro-quinolin-6-yl)-acetic acid as a brown solid (0.23 g),

Assay Found: C 65.2; H 6.95; N 6.9% C₂₂ H₂₇ ClN₂ O₃.0.1H₂ O requires C65.3; H 6.8; N 6.9%

n.m.r. (DMSO-d₆): d values include 0.99 (d, 3H), 3.30 (s, 2H), 4.61 (t,1H), 6.50 (d, 1H), 6.73 (s, 1H), 6.81 (dd, 1H), 7.37 (broad s, 1H).

from[1-(2-{tert-Butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanoxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propyl)-1,2,3,4-tetrahydro-quinolin-6-yl]-aceticacid methyl ester (0.51 g).

EXAMPLE 3

(4-{2R-[2-(3-Chloro-phenyl)-2R-hydroxyl-ethylamino]-propylamino}-2-methyl-phenyl)-aceticacid as a colourless solid (0.08 g),

Assay Found: C 55.5; H 6.7; N 8.7% C₂₀ H₂₅ ClN₂ O₃.1 H₂ O.0.75NH₄ Clrequires C 55.2; H 6.95; N 8.85%

n.m.r. (DMSO-d₆): d values include 1.06 (d, 3H), 2.11 (s, 3H), 4.62 (t,1H), 6.34 (t, 2H), 6.87 (d, 1H), 7.22-7.50 (m, 4H).

from[4-(2R-{tert-Butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanoxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propylamino)-2-methyl-phenyl]-aceticacid methyl ester (0.2 g).

EXAMPLE 4

(4-{2R-[2-(3-Chloro-phenyl)-2R-hydroxyl-ethylamino]-propylamino}-3-methyl-phenyl)-aceticacid as a colourless solid (0.11 g),

Assay Found: C 45.45; H 6.95; N 11.1% C₂₀ H₂₅ ClN₂ O₃.2.6H₂ O.3.3NH₄ Clrequires C 45.3; H 6.9; N 11.2%

n.m.r. (DMSO-d₆): d values include 1.10 (d, 3H), 2.02 (s, 3H), 4.71 (t,1H), 6.49 (d, 1 H), 6.88 (broad d, 2H), 7.21-7.49 (m, 4H).

from[4-(2R-{tert-butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanoxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propylamino)-3-methyl-phenyl]-aceticacid methyl ester (0.24 g).

EXAMPLE 5

(4-{2R-[2-(3-Chloro-phenyl)-2R-hydroxyl-ethylamino]-propylamino}-2-fluoro-phenyl)-aceticacid as a colourless solid (0.08 g),

Assay Found: C 49.1; H 5.9; N 9.65% C₁₉ H₂₂ ClFN₂ O₃.0.8H₂ O.1.3NH₄ Clrequires C 49.1; H 6.25; N 9.9%

n.m.r. (DMSO-d₆): d values include 1.08 (d, 3H), 4.67 (t, 1H), 6.26-6.42(m, 2H), 6.96 (t, 1H), 7.23-7.49 (m, 4H).

from[4-(2R-{tert-butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanoxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propylamino)-2-fluoro-phenyl]-aceticacid methyl ester (0.24 g).

EXAMPLE 6

(4-{2R-[2-(3-Chloro-phenyl)-2R-hydroxyl-ethylamino]-propylamino}-3-fluoro-phenyl)-aceticacid as a pale brown foam (0.36 g),

Assay Found: C 56.8; H 5.8; N 7.3% C₁₉ H₂₂ ClFN₂ O₃.H₂ O requires C57.2; H 6.1; N 7.0%

n.m.r. (DMSO-d₆): d values include 1.08 (d, 3H), 4.71 (t, 1H), 6.66 (t,1H), 6.79-7.02 (q, 2H), 7.21-7.51 (m, 4H).

from[4-(2R-{tert-butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanoxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propylamino)-3-fluoro-phenyl]-aceticacid methyl ester (0.50 g).

EXAMPLE 7

(4-{2R-[2-(3-Chloro-phenyl)-2R-hydroxyl-ethylamino]-propylamino}-2,3-difluoro-phenyl)-aceticacid as a colourless solid (0.20 g),

Assay Found: C 37.2; H 6.4; N 11.4% C₁₉ H₂₁ ClF₂ N₂ O₃.3H₂ O.3NH₄ Clrequires C 37.1; H 6.2; N 11.9%

n.m.r. (DMSO-d₆): d values include 1.14 (d, 3H), 5.02 (t, 1H), 6.67(broad t, 1H), 6.90 (t, 1H), 7.28-7.60 (m, 4H)

from[4-(2R-{tert-butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanoxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propylamino)-2,3-difluoro-phenyl]-aceticacid methyl ester (0.44 g).

EXAMPLE 8

(5-Chloro-4-{2R-[2-(3-chloro-phenyl)-2R-hydroxy-ethylaminol-propylamino}-2-methoxy-phenyl)-aceticacid dihydrochloride as a colourless solid (0.006 g)

n.m.r. (DMSO-d₆): d values include 1.04 (s, 3H), 3.68 (s, 3H), 4.65 (t,1H), 7.00 (s, 1H), 7.21-7.35 (m, 4H), 7.39 (s, 1H)

from(5-chloro-[4-(2R-{tert-butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanoxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propylamino)-2-methoxy-phenyl])-aceticacid methyl ester (0.037 g).

EXAMPLE 9

(4-{2-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-ethylamino}-2-methyl-phenyl)-aceticacid as a colourless solid (0.287 g)

C₁₉ H₂₄ Cl N₂ O₃ : MH⁺ 363.1470 (error 1.6 ppm)

n.m.r. (DMSO-d₆): d values include 2.11 (s, 3H), 4.65 (m, 1H), 6.32 (dd,1H), 6.34 (d, 1H), 6.84 (d, 1H), 7.40 (broad s, 1H)

from4-(2-(tert-Butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-ethylamino)-2-methyl-phenyl]-aceticacid methyl ester (0.549 g)

EXAMPLE 10

(4-{2-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-ethylamino}-2-trifluoromethyl-phenyl)-aceticacid as a yellow solid (0.028 g)

C₁₉ H₂₁ Cl F₃ N₂ O₃ : MH⁺ 417.1208 (error 3.6 ppm)

n.m.r. (DMSO-d₆): d values include 3.54 (s, 2H), 4.68 (m, 1H), 6.76 (dd,1H), 6.84 (d, 1H), 7.15 (d, 1H), 7.41 (broad s, 1H)

from[4-(2-{tert-Butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-ethylamino)-2-trifluoromethyl-phenyl]-aceticacid methyl ester (0.18 g).

EXAMPLE 11

(4-{2-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-ethylamino}-2,3-difluoro-phenyl)-aceticacid from[4-(2-{tert-Butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-ethylamino)-2,3-difluoro-phenyl]-aceticacid methyl ester.

EXAMPLE 12

(4-{2R-[2-(3-Chloro-phenyl)-2R-hydroxyl-ethylamino]-amino}-2,3,6-trifluoro-phenyl)-aceticacid from[4-(2R-{tert-Butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propylamino)-2,3,6-trifluoro-phenyl]-aceticacid methyl ester.

EXAMPLE 13

[2-Chloro-4-(2R-{[2-(3-chloro-phenyl)-2R-hydroxy-ethyl]-amino}-propylamino)-phenyl]-aceticacid (15 mg) as a white solid.

¹ H NMR (300 mHz, DMSO-d₆) d 7.4(s,₁ H), 7.3 (m,3H), 7.0 (d, 1H), 6.6(s, 1H), 6.5 (d, 1H), 5.8 (bs, 1H), 4.7 (m,1 H), 1.1 (d, 3H).

HPLC analysis (C18 column) 1mL/min, 20-50% acetonitrile/water(0.1% TFA)T_(R) =17.5 min

from[4-(2R-{tert-Butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanyioxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propylamino)-2-chloro-phenyl]-aceticacid methyl ester.

EXAMPLE 14

[5-Chloro-4-(2R-{[2-(3-chloro-phenyl)-2R-hydroxy-ethyl]-amino}-propylamino)-2-methyl-phenyl]-aceticacid methyl ester as a white solid (110 mg)

Analysis: Calc. for C₂₀ H₂₄ N₂ O₃ Cl₂ (0.25 H₂ O): C, 56.55, H, 5.69, N,6.59% Found: C, 56.49, H, 5.99, N, 6.56% C₂₀ H₂₄ N₂ O₃ Cl₂ : MH⁺ 411

from[4-(2R-{tert-Butoxycarbonyl-[2R-(tert-butyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propylamino)-5-chloro-2-methyl-phenyl]-aceticacid methyl ester.

EXAMPLE 15

(4-{2R-[2-(3-Chloro-phenyl)-2R-hydroxy-ethylamino]-propylamino}-2,6-difluoro-phenoxy)-aceticacid as an off-white solid (46 mg)

mp 105° C. (dec)

n.m.r (CDCl3): d 1.34 (d, 3H), 3.05-3.57 (m, 5H), 4.30 (s, 2H), 5.00(dd, 1H), 6.20-6.30 (m, 2H), 7.28-7.40 (m, 3H), 7.47 (s, 1H).

from(4-[2R-{tert-butoxycarbonyl-[2R-(tertbutyl-dimethyl-silanyloxy)-2-(3-chloro-phenyl)-ethyl]-amino}-propylamino)-2,6-difluoro-phenyoxy]-aceticacid methyl ester (451 mg)

EXAMPLE 16

(4-{2R-[2-(3-Chloro-phenyl)-2R-hydroxy-ethylamino]-propylamino}-3-trifluoromethyl-phenyl)-aceticacid

A solution of(4-{2R-[2-(3-chloro-phenyl)-2R-hydroxy-ethylamino]-propylamino}-3-trifluoromethyl-phenyl)-aceticacid methyl ester (75.1 mg) in tetrahydrofuran (4 mL) and 6N aqueoushydrochloric acid (4 mL) was stirred at ambient temperature for 18 h.The residue was concentrated and lyophilized to afford a residue whichwas purified by silica gel chromatography eluting withchloroform:methanol:concentrated ammonium hydroxide (30:15:1) to affordafter concentration of the relevant fractions the title compound (58.9mg) as a white solid.

mp 161-163° C.

Assay: Found: C 54.5, H. 5.2, N, 6.1% C₂₀ H₂₂ Cl₁ F₃ N₂ O₃.0.5H₂ Orequires C 54.6, H. 5.3, N. 6.4%

EXAMPLE 17

(4-{2R-[2-(3-Chloro-Phenyl)-2R-hydroxy-ethylamino]-propylamino}-2-triftuoromethyl-phenyl)-aceticacid

A solution of(4-{2R-[2-(3-chloro-phenyl)-2R-hydroxy-ethylamino]-propylamino}-2-trifluoromethyl-phenyl)-aceticacid methyl ester (290 mg) in tetrahydrofuran (4 mL) and 6N aqueoushydrochloric acid (4 mL) was stirred at ambient temperature for 18 h.The residue was concentrated and lyophilized to afford a residue whichwas purified by silica gel chromatography eluting withchloroform:methanol:concentrated ammonium hydroxide (30:15:1) to affordafter concentration of the relevant fractions the title compound (122mg) as a white solid.

C₂₀ H₂₂ Cl₁ F₃ N₂ O₃ MH⁺ 431.134 (error 2.2 ppm)

n.m.r (CDCl₃): d values include 1.085 (d, 3H), 2.85 (m, 2H), 2.885-3.12(m, 3H), 3.545 (s, 2H) 4.705 (m, 1H), 6.045 (bt, 1H), 6.765 (dd, 1H),6.855 (d, 1H), 7.265-7.4 (m, 3H).

TABLETS FOR ORAL ADMINISTRATION

Tablets may be prepared by the normal methods such as direct compressionor wet granulation.

The tablets may be film coated with suitable film forming materials,such as hydroxypropyl methylcellulose, using standard techniques.Alternatively the tablets may be sugar coated.

    ______________________________________                                        Direct Compression Tablet                                                                              mg/tablet                                            ______________________________________                                        (i)      Active Ingredient      4.688                                                     Calcium Hydrogen Phosphate BP*                                                                       83.06                                                  Croscarmellose Sodium NF                                                                                    1.8                                             Magnesium Stearate BP                                                                                         0.                                            Compression weight                                                                                               90.0                           ______________________________________                                         *of a grade suitable for direct compression.                             

The active ingredient is passed through a 60 mesh sieve, blended withthe calcium hydrogen phosphate, croscarmellose sodium and magnesiumstearate. The resultant mix is compressed into tablets using a ManestyF3 tablet machine fitted with 5.5 mm, flat bevelled edge punches.

    ______________________________________                                                            mg/tablet                                                 ______________________________________                                        (ii)      Active Ingredient.                                                                                              0.31                                            Anhydrous Lactose USNF                                                                                  131.99                                              Pregelatinised Starch USNF                                                                            7.0                                                   Magnesium Stearate BP                                                                                      0.7                                              Compression weight                                                                                          140.0                             ______________________________________                                    

The active ingredient is passed through a 60 mesh sieve, and blendedwith the lactose, pregelatinised starch and magnesium stearate. Theresultant mix is compressed into tablets using a Manesty F3 tabletmachine fitted with 7.5 mm normal concave punches.

SYRUP

This may be either a sucrose or sucrose free presentation.

    ______________________________________                                        A.        Sucrose Syrup    mg/5 ml dose                                       ______________________________________                                                Active Ingredient                                                                                2.5                                                        Sucrose BP                 2750.0                                             Glycerine BP                                                                                             500.0                                              Buffer                                                                        Flavour                                                                       Colour                  as required                                           Preservative                                                                  Purified Water BP                                                                                          5.0 m                                    ______________________________________                                    

The active ingredient, buffer, flavour, colour and preservative aredissolved in some of the water and the glycerine is added. The remainderof the water is heated to dissolve the sucrose and is then cooled. Thetwo solutions are combined, adjusted to volume and mixed. The syrup isclarified by filtration.

    ______________________________________                                        B.     Sucrose-free Syrup    mg/5 ml dose                                     ______________________________________                                        Active Ingredient       2.5                                                   Hydroxypropylmethylcellulose USP                                              (viscosity type 4000)                22.5                                     Buffer                                                                        Flavour                                                                       Colour                        as required                                     Preservative                                                                  Sweetener                                                                     Purified Water BP to         5.0 ml                                           ______________________________________                                    

The hydroxypropylmethylcellulose is dispersed in hot water, cooled andthen mixed with an aqueous solution containing the active ingredient andthe other components of the formulation. The resultant solution isadjusted to volume and mixed. The syrup is clarified by filtration.

INJECTION FOR INTRAVENOUS ADMINISTRATION

    ______________________________________                                                                μg/ml                                              ______________________________________                                        (i)    Active Ingredient                       800                                      Dilute Hydrochloric Acid BP to pH 3.5                                          Sodium Chloride Injection BP to l ml                               ______________________________________                                    

The active ingredient is dissolved in a suitable volume of SodiumChloride Injection BP, the pH of the resultant solution is adjusted topH3.5 with dilute hydrochloric acid BP then the solution is made tovolume with sodium chloride injection BP and thoroughly mixed. Thesolution is filled into Type I clear glass 5 ml ampoules which aresealed under a headspace of air, by fusion of the glass then sterilisedby autoclaving at 120⁰ for not less than 15 minutes.

    ______________________________________                                                               μg/ml                                               ______________________________________                                        (ii)      Active ingredient                 56.2                                             Sodium Chloride BP                                                                                      as required                                         Water for Injection BP to                                                                            1.0 ml                                  ______________________________________                                    

Sodium chloride may be added to adjust the tonicity of the solution andthe pH may be adjusted, using acid or alkali, to that of optimumstability and/or facilitate solution of the active ingredient.Alternatively, suitable buffer salts may be used.

The solution is prepared, clarified and filled into appropriate sizeampoules sealed by fusion of the glass. The injection is sterilised byheating in an autoclave using one of the acceptable cycles.Alternatively, the solution may be sterilised by filtration and filledinto sterile ampoules under aseptic conditions. The solution may bepacked under an inert atmosphere of nitrogen or other suitable gas.

SUPPOSITORY FOR RECTAL ADMINISTRATION

    ______________________________________                                        Active ingredient     49.0   mg                                               Witepsol* H15 to            1.0                                                                            g                                                ______________________________________                                         *a proprietary grade of Adeps Solidus Ph.Eur.                            

A suspension of the active ingredient in molten Witepsol is prepared andfilled using suitable machinery, into 1g size suppository moulds.

The compounds of Examples 7, 8 and 15 were tested forbeta-3-adrenoceptor activity using above described Method I with thefollowing results:

    ______________________________________                                                  EPMR                                                                Test Method  Ex 7        Ex 8   Ex l5                                         ______________________________________                                        Method 1     0.2         0.4     6.0                                          ______________________________________                                    

The protective effect of the compound of Example 7 was measured asdescribed in above Method 2 and an ED₅₀ of 0.003 mg/kg was obtained.

We claim:
 1. A compound of the general formula (I): ##STR8## wherein R¹represents an aryl group optionally substituted by one or moresubstituents selected from halogen, hydroxy, C₁₋₆ alkoxy, C₁₋₆ alkyl,nitro, cyano, hydroxymethyl and trifluoromethyl;R² represents hydrogenor C₁₋₆ alkyl; R³ represents a group A ##STR9## where the ring issubstituted by one to four further substituents selected from C₁₋₆alkyl, halogen, trifluoromethyl, and C₁₋₆ alkoxy; or R³ represents agroup B ##STR10## where the aromatic ring is optionally substituted byup to three further substituents selected from C₁₋₆ alkyl, halogen,trifluoromethyl, and C₁₋₆ alkoxy; R⁴ represents hydrogen, or C₁₋₆ alkyl;R⁵ represents ZCH₂ CO₂ H wherein Z represents a bond, or O; Y represents(CH₂), where n is 1-3; and physiologically acceptable derivativesthereof.
 2. A compound as claimed in claim 1 wherein R¹ represents aphenyl group substituted by a chlorine atom located in the metaposition.
 3. A compound as claimed in claim 1 wherein R² is methyl or H.4. A compound as claimed in claim 1 where R³ is group A substituted byone or more substituents selected from halogen, methyl, trifluoromethyl,and methoxy.
 5. A compound as claimed in claim 1 where R³ is group B, nis 1 or 2, and the aromatic ring is unsubstituted.
 6. A compound asclaimed in claim 1 where R⁴ is hydrogen or methyl.
 7. A compound asclaimed in claim 1 where R¹ represents phenyl substituted by a chlorineatom located in the meta position, R² represents hydrogen or methyl, R³represents a group A and is substituted by one or more groups selectedfrom halogen, methyl, trifluoromethyl, and methoxy, R⁴ representshydrogen or methyl, R⁵ represents CH₂ CO₂ H, and physiologicallyacceptable derivatives thereof.
 8. A compound selected from the groupconsistingof(1-{2-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-propyl}-2,3-dihydro-1H-indol-5-yl)-aceticacid;(1-{2-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-propyl}-1,2,3,4-tetrahydro-quinolin-6-yl)-aceticacid;(4-{2R-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-propylamino}-2-methyl-phenyl)-aceticacid;(4-{2R-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-propylamino}-3-methyl-phenyl)-aceticacid;(4-{2R-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-propylamino}-2-fluoro-phenyl)-aceticacid;(4-{2R-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-propylamino}-3-fluoro-phenyl)-aceticacid;(4-{2R-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-propylamino}-2,3-difluoro-phenyl)-aceticacid;(5-Chloro-4-{2R-[2-(3-chloro-phenyl)-2R-hydroxy-ethylamino]-propylamino}-2-methoxy-phenyl)-aceticacid;(4-{2-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-ethylamino}-2-methyl-phenyl)-aceticacid;(4-{2-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-ethylamino}-2,3-difluoro-phenyl)-aceticacid;(4-{2-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-ethylamino}-2-trifluoromethyl-phenyl)-aceticacid;(4-{2R-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-propylamino}-2,6-difluoro-phenyl)-aceticacid;(4-{2-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-ethylamino}-2,3,6-trifluoro-phenyl)-aceticacid;(4-{2-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-propylamino}-2-trifluoromethyl-phenyl)-aceticacid;(4-{2R-[2-(3-Chloro-phenyl)-2R-hydroxyl-ethylamino]-propylamino}-2,3,6-trifluoro-phenyl)-aceticacid;[2-Chloro-4(2-{[2-(3-chloro-phenyl)-2R-hydroxy-ethyl]-amino}-propylamino)-phenyl]-aceticacid;[5-Chloro-4-(2R-{[2-(3-chloro-phenyl)-2R-hydroxy-ethyl]-amino}-propylamino)-2-methyl-phenyl]-aceticacid;(4-{2R-[2-(3-Chloro-phenyl)-2R-hydroxy-ethylamino]-propylamino}-2,6-difluoro-phenoxy)-aceticacid and;(4-{2R-[2-(3-Chloro-phenyl)-2R-hydroxy-ethylamino]-propylamino}-3-trifluoromethyl-phenyl)-aceticacid;or a physiologically acceptable derivative thereof. 9.(4-{2R-[2-(3-chloro-phenyl)-2R-hydroxyl-ethylamino]-propylamino}-2,3-difluoro-phenyl)-aceticacid;or a physiologically acceptable derivative thereof.
 10. A method oftreatment of a mammal suffering from a condition susceptible ofamelioration by an atypical beta-adrenoceptor agonist comprisingadministration of an effective amount of a compound according to claim 1or a physiologically acceptable derivative thereof.
 11. A pharmaceuticalcomposition comprising a compound according to claim 1 or aphysiologically acceptable derivative thereof together with one or morepharmaceutically acceptable carriers.
 12. A pharmaceutical compositionwhich comprises a compound according to claim 1 and a non-steroidalanti-inflammatory drug, together with one or more pharmaceuticallyacceptable carriers.
 13. A process for preparing a compound of formula(I) as claimed in claim 1, or a physiologically acceptable derivativethereof which comprises:(A), compounds of formula (I) may be prepared byreaction of a compound of formula (Ia) ##STR11## wherein R¹, R², and,are as defined as for formula (I), R³ represents R³ where the acidicgroup is protected by an alkyl ester, and R^(a) and R^(b) are protectinggroups, by deprotection of the protecting groups or (B), compounds offormula (I) may be prepared by reaction of a compound of formula (II)with a compound of formula (III): ##STR12## wherein R¹, R², R³, R^(a),and R^(b) are as defined above, in the presence of a reducing agent,followed by removal of the protecting groups.